Receptor-mediated vectorial transcytosis of epidermal growth factor by Madin-Darby canine kidney cells.

Maratos–Flier, Eleftheria; Yang, Chiou-Ying; Verdin, Eric; King, George L.

doi:10.1083/jcb.105.4.1595

Cited by 126 publications

(46 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…While both apical to basolateral and basolateral to apical transcytosis are also blocked at 17°C (Maratos-Flier et al, 1987;Breitfeld et al, 1989;Hunziker and Mellman, 1989), the temperature sensitive step(s) has been unknown. Since at 17°C, dIgA-containing vesicles accumulated in the apical cytoplasm, neither the formation of transcytotic vesicles from basolateral endosomes nor vesicle translocation was affected.…”

Section: Discussionmentioning

confidence: 99%

Differential microtubule requirements for transcytosis in MDCK cells.

1990

View full text Add to dashboard Cite

Given the role of microtubules in directing the transport of many intracellular organelies, we investigated whether intact microtubules were also required for transcytosis across epithelia. Using polarized MMCK cells expressing receptors for the Fc domain of IgG (FcRH-B2) or polymeric immunoglobulin (plg-R), we examined the involvement of microtubules in apical to basolateral and basolateral to apical transcytosis, respectively. While depolymerization of microtubules with nocodozole had no effect on apical to basolateral transcytosis via FcR, basolateral to apical transcytosis of dimeric IgA via plg-R was alnost completely blocked. Inhibition due to nocodozole was selective for basolateral to apical transcytosis, since neither endocytosis nor receptor recycling was significantly affected at either plasma membrane domain. As shown by confocal microscopy, the block in transcytosis was due to the inability of MDCK cells to translocate IgA-containing vesicles from the basolateral to the apical cytoplasm in the absence of an intact microtubule network. The nocodazole sensitive step could be partially by-passed, however, by allowing cells to internalize IgA at 17°C prior to nocodazole treatment. Although incubation at 17°C blocked release of IgA into the apical medium, it did not prevent translocation of IgA-containing vesicles to the apical cytoplasm. Thus, receptor-mediated transcytosis in opposite directions exhibits distinct requirements for microtubules, a feature which reflects the spatial organization of MMCK cells.

show abstract

Section: Discussionmentioning

confidence: 99%

Differential microtubule requirements for transcytosis in MDCK cells.

1990

View full text Add to dashboard Cite

show abstract

“…The organization of the plasma membrane of polarized epithelial cells is complex, with functionally and biochemically distinct apical and basolateral domains separated by tight junctions that block intercellular diffusion of ions and macromolecules (reviewed in reference 53). EGFRs are located primarily on the basolateral surface of most polarized epithelial cells, including human airway tracheal epithelial cells (unpublished results) and canine kidney tubule epithelial MDCK cells (45). Moreover, a fraction of ligand-bound EGFRs are transcytosed to the apical surface in MDCK cells (45).…”

Section: Discussionmentioning

confidence: 99%

Adenovirus E3 protein causes constitutively internalized epidermal growth factor receptors to accumulate in a prelysosomal compartment, resulting in enhanced degradation.

Hoffman¹,

Carlin²

1994

Mol. Cell. Biol.

View full text Add to dashboard Cite

We have previously identified and characterized an integral membrane protein coded for by the early transcription region 3 (E3) of human group C adenoviruses that down-regulates the epidermal growth factor receptor (EGFR). The goal of this study was to characterize the early receptor trafficking events leading to enhanced EGFR degradation in adenovirus-infected cells. Specifically, we wished to determine whether adenovirus increases the rate of EGFR internalization or alters the subcellular compartmentalization of internalized EGFRs. Once the optimal time for measuring early trafficking events was determined, surface EGFRs were labeled with a cleavable biotin reagent to measure internalization rates and with a receptorspecific monoclonal antibody (MAb) conjugated to colloidal gold for intracellular localization studies. We first showed that the rate of EGFR internalization in adenovirus-infected cells is indistinguishable from the constitutive internalization rate for unoccupied EGFRs. The possibility that the E3 protein can affect trafficking of EGFRs internalized at a low constitutive rate was further supported by studies showing that adenovirus-mediated down-regulation occurs independently of EGFR oligomerization and intrinsic EGFR tyrosine kinase activity, which are required for efficient ligand-induced internalization. Other tyrosine kinases inhibited by genistein are also not required for adenovirus-induced down-regulation. When the intracellular localization of EGFRs during adenovirus-mediated down-regulation was examined by electron microscopy, there was a threefold increase in the number of EGFRs localized to multivesicular bodies. The multivesicular body has been proposed to be important for regulating intracellular membrane protein sorting, since trafficking patterns for receptors that recycle and receptors that are degraded diverge in this organelle. These data therefore suggest that adenovirus may enhance EGFR degradation by causing constitutively internalized EGFRs to accumulate in a prelysosomal compartment. This is the first example of a mechanism that efficiently down-regulates EGFR without significantly increasing the rate of internalization or that does not require EGFR tyrosine kinase activity. Since viral proteins often mimic or modify a host counterpart, this suggests that there are normal physiological conditions when receptor destruction without tyrosine signalling is beneficial.Epidermal growth factor (EGF) and transforming growth factor a are secreted polypeptides that regulate proliferation and differentiation of a variety of cell types (reviewed in reference 10). Their effects are mediated through binding to specific cell surface receptors belonging to a family of evolutionarily conserved proteins with intrinsic tyrosine kinase activity (reviewed in references 9 and 58). The EGF and transforming growth factor a receptor (EGFR) has a large extracellular glycosylated ligand-binding domain, a single hydrophobic membrane-spanning domain, and a cytoplasmic domain containing a kinase cataly...

show abstract

“…These surfaces are separated by tight junctions (Martinex-Palomo et al, 1980) and are functionally different (Louvard, 1980;Misfeldt et al, 1976;Pesonen & Simons, 1983). The cells are capable of transcellular transport across the cytoplasm of a variety of ligands, including viral proteins (Pesonen & Simons, 1983), immunoglobulins (Mostov & Deitcher, 1986) and epidermal growth factor (EGF) (Maratos-Flier et al, 1987). In addition, entry of certain viruses is polarized to either the apical (Clayson & Compans, 1988) or basal (Basak & Compans, 1989) surface.…”

Section: Introductionmentioning

confidence: 99%